Figure 2
PC is required for virus-triggered expression of IFN and ISG.
(A) A549 cells were transfected with PC-specific siRNA or nonsense control siRNA for 48 h and PC mRNA levels were determined by qPCR. **p < 0.01 (one-way ANOVA). (B) A549 cells were transfected with different PC-specific siRNA and control siRNA. Fourty-eight hours after transfection, MTS assays were performed using a MTS cell viability kit (Promega) according to the manufacturer’s instructions. Cell viability was then calculated. no sign., no significant difference (one-way ANOVA). (C) A549 cells were co-transfected with the indicated PC-specific siRNA or nonsense control siRNA and IFN-β or IFN-λ1 luciferase reporter plasmids for 24 h. pRL-TK was cotransfected as an internal control in the luciferase assay. Luciferase assays were performed after cells were infected with SeV (MOI = 1) for 12 h. **p < 0.01 (one-way ANOVA). (D) PC-KD and Scr.-KD cells were collected to assess PC mRNA and protein levels by qPCR and western blot, respectively. **p < 0.01 (one-way ANOVA). (E) The same numbers of PC-KD and Scr.-KD cells were infected with SeV (MOI = 1) for 6 h. IFN mRNA levels were measured by qPCR. *p < 0.05, **p < 0.01 (one-way ANOVA). (F) PC-KD and Scr.-KD cells were infected with SeV (MOI = 1) for 12 h. PKR and Mx1 mRNA and protein levels were determined by qPCR and western blot, respectively. *p < 0.05 (one-way ANOVA). All graphs represent means standard deviations for 3 experiments.
